Living organisms have evolved multienzyme complexes to achieve efficient and spatially ordered metabolic reactions. Recently, liquid-liquid phase separation (LLPS) has emerged as a fundamental organizational principle underlying these natural networks, providing a versatile platform for constructing artificial multienzyme catalytic systems. Owing to their reversible self-assembly and programmable nature, LLPS condensates can modulate enzyme distribution, mass transfer, and the reaction microenvironment, thereby significantly enhancing catalytic efficiency and cascade reaction throughput. This review systematically summarizes recent advances in in vitro LLPS-based multienzyme catalytic systems. We introduce the fundamental phase separation behaviors of multienzyme condensates and their molecular interaction mechanisms. Then, diverse construction strategies are discussed based on distinct intermolecular forces and their applications in cofactor recycling, biosynthesis, biodegradation, and artificial organelle fabrication are highlighted. Furthermore, we dissect the core mechanisms of catalytic enhancement, including local molecular enrichment, microenvironment modulation, and spatial confinement. Finally, we discuss current limitations and offer future perspectives for the rational design of advanced, high-efficiency multienzyme biocatalysts.
Biomolecular condensates, formed by proteins, nucleic acids, and other macromolecules through liquid-liquid phase separation, enable molecular enrichment and reaction regulation within membraneless microenvironments. As their physical principles have become better understood, condensates are increasingly being incorpor...
Confined enzyme catalysis is a promising approach for pharmaceutical synthesis because it can combine high selectivity, catalytic efficiency and mild reaction conditions. This review examines recent advances in multiscale enzyme confinement for pharmaceutical applications, with emphasis on atomic-scale active-site regu...
Enzymatic reactions in biomolecular condensates are often assumed to be regulated through local enrichment of reactants. However, condensates also reshape molecular transport and reaction kinetics, making it unclear how phase separation controls catalysis in living cells. Here, we develop a quantitative theory of biomo...
Gaetano Granatelli, Samuel S Gomez, Sudarshana Laha et al.· bioRxiv· 0 citations
Biomolecules are nature's catalysts and recognition elements, yet their practical use is limited by instability and lack of reusability. Crystalline porous organic frameworks (POFs) have emerged as protective hosts, but the field is now shifting from passive encapsulation toward engineering multi-component synergy. Thi...
Jiaxin Cui, Xin Liu, Jing-Ting He et al.· Chemical Society Reviews· 0 citations
Nucleic acid condensates are emerging as programmable soft materials capable of organizing and regulating biochemical reactions through sequence-directed interactions. Inspired by biological membrane-less organelles formed by liquid-liquid phase separation, synthetic DNA and RNA condensates can selectively concentrate,...
Richard J. Archer, Hirotake Udono, Masahiro Takinoue· Current Opinion in Structura...· 0 citations
Heterojunction nanozymes offer unique advantages, including interface charge regulation, multivalent-state cycling, defect engineering, and enhanced field responses. These merits overcome the bottlenecks of single-component nanozymes, such as low catalytic activity, poor stability, and limited functionality. They hav...
